Potential Biotechnological Tool Research Articles

Uptake of spherical nucleic acid (SNA) in Ochromonas danica: A new potential biotechnological tool

The development of a nanomaterial-based platform for application as an in vivo sensor or genome-editing tool is of great importance to microalgal biotechnology. In this study, gold nanoparticles (AuNPs) were functionalized with DNA to form spherical nucleic acid (SNA) through a pH-assisted method. The fabricated SNA readily entered Ochromonas danica, a microalga species that is suitable to study the endocytosis mechanisms, after the cells were simply incubated with SNA solution for 5 h. The transmission electron microscopy (TEM) images demonstrated the internalization of SNA within both the cytoplasm and vacuole, indicating their intracellular presence and distribution. The uptake of SNA was significantly inhibited by the clathrin-mediated endocytosis inhibitor (chlorpromazine, CPZ) and caveolae-mediated endocytosis inhibitor (methyl-β-cyclodextrin, MβCD), suggesting that SNA was taken up by O. danica cells mainly through the clathrin- and caveolae-dependent pathways. The toxic effects of SNA on O. danica were negligible since the growth and photosynthesis of treated cells were comparable to those of untreated cells (control). As a proof-of-concept, the SNA platform was used as an intracellular sensor for microalgae by using an aptamer-based SNA (i.e., aptamer nano-flare). The aptamer nano-flare fluorescently imaged the intracellular ATP, showing the high potential for SNA applications. This study will contribute to the application of nanotechnology to improve microalgal biotechnology.

Genetic basis for the biodegradation of a polyether-polyurethane-acrylic copolymer by a landfill microbial community inferred by metagenomic deconvolution analysis

Plastic accumulation in the world amounts to approximately 8300 million tons. Polyurethanes (PU) account for 7.7 % of total plastics production worldwide, and their diverse chemical composition makes them highly recalcitrant to biodegradation. Several works have reported polyurethane-degrading microbial communities. However, it is still necessary to learn more about the chemical, biochemical, and genetic bases linked to the polyurethanolytic phenotype and the microbial taxonomic determinants responsible for metabolizing the PU polymer and its associated chemical additives. To shed light on this problem, we applied physical, chemical, biochemical, metagenomic, and bioinformatic analyses to explore the biodegradation capability and related biochemical and genetic determinants of the BP6 microbial community that can grow in PolyLack, a commercial coating containing a polyether polyurethane acrylate (PE-PU-A) copolymer and several additives, as sole carbon source. We observed complete additives (isopropanol, N-methyl-2-pyrrolidone, 2-butoxyethanol, alkyl glycol ethers) biodegradation and the appearance of released polymer components (toluene diisocyanate (TDI) and methylene diphenyl diisocyanate (MDI) derivatives), and multiple degradation products since early cultivation times. The Hi-C metagenomic analysis identified a complex microbiome with 35 deconvolved Metagenome-Assembled Genomes (MAGs) – several new species – and biodegradation markers that suggest the coexistence of hydrolytic, oxidative, and reductive metabolic strategies for degrading the additives and the PU copolymer. This work also provides evidence of the metabolic capability the BP6 community has for biodegrading polyether polyurethane foams. Based on these analyses, we propose a novel metabolic pathway for 4,4′-methylenedianiline (MDA), an initial biodegradation intermediate of MDI-based PU, encoded in the complex BP6 community metagenome and suggest that this community is a potential biotechnological tool for PU bio-recycling.

Gene Silencing of laccase 1 Induced by Double-Stranded RNA in Callosobruchus maculatus (Fabricius 1775) (Coleoptera: Chrysomelidae) Suggests RNAi as a Potential New Biotechnological Tool for Bruchid’s Control

Bruchids are the most important pests of leguminous seeds in the world. In this study, the focus was done on Callosobruchus maculatus, a serious pest of Vigna unguiculata seeds. As no efficient control methods preventing collateral effects on beneficials currently exist, this study investigated whether RNA interference (RNAi) could provide a new biotechnological and selective tool for bruchids control. Three principal objectives were followed including (i) the identification of all RNAi machinery core components and a key protein to silence in C. maculatus genome (c.f., dicer-2, argonaute-2, R2D2, and laccase 1), (ii) the identification of suitable reference gene for RT-qPCR analyses, and (iii) the micro-injection of dsRNA coding for laccase 1 to adults of C. maculatus to assess gene expression levels by RT-qPCR and potentially related mortalities. Phylogenetical analyses performed from transcriptomic information successfully identified all necessary proteins in the RNAi mechanism and also the open reading frame of laccase 1 in C. maculatus. A new reference gene was identified (i.e., alpha-tubuline 1) and coupled with glutiathone S transferase for RT-qPCR analyses. Double-stranded RNAs coding for laccase 1 and green fluorescent protein (control) were produced and 400 ng of each dsRNA were micro-injected into C. maculatus adults. RT-qPCR analyses revealed a stable significant decrease in laccase 1 expression in about 80% of adults treated with laccase 1 dsRNA after three days post-injection. No significant mortalities were observed which is probably related to the non-exposure of adults to anti-nutritional factors that are usually regulated by laccase. Further research should focus either on the feeding larval stage which is directly exposed to anti-nutritional factors, or on other target genes to induce dead phenotypes. This study is the first gene silencing report on a bruchid species and supports RNAi as a potential future method of control.

Genome sequence of Pseudomonas aeruginosa PA1-Petro—A role model of environmental adaptation and a potential biotechnological tool

Pseudomonas aeruginosa is a ubiquitous microorganism, capable of colonizing a wide range of habitats due to its metabolic versatility and wide adaptability to different conditions. Industrial and environmental research involving petroleum microbiology play a pivotal role in controlling many technical, operational, and environmental issues. P. aeruginosa PA1-Petro strain was isolated from oil production water in Northeastern Brazil. Herein we report the genomic sequencing and annotation of PA1-Petro, and a comparative genomics study against two widely used reference P. aeruginosa strains (PAO1 and PA14). PA1-Petro has a genome of 6,893,650 bp, the largest among the three analyzed in this study, with a 65.87% GC content. The analyzes resulted in a wide repertoire of 544 unique genes in PA1-Petro, and the highest copy numbers of common genes among the three strains (PA1-Petro, PAO1 and PA14). Unique sequences are hypothetical proteins, prophage sequences, mobile genetic elements, transcriptional regulators, metal resistance genes to copper, tellurium and arsenic, type IE CRISPR-Cas, Type VI Secretion System (T6SS)-associated proteins, and a toxin-antitoxin system. Taken together, these results provide intriguing insights on adaptive evolution within PA1-Petro genome, adding unprecedented information to the species’ plasticity and ubiquitous characteristics.

Screening for a more sustainable solution for decolorization of dyes and textile effluents using Candida and Yarrowia spp.

Dyed effluents from textile industry are toxic and difficult to treat by conventional methods and biotechnological approaches are generally considered more environmentally friendly. In this work, yeast strains Candida parapsilosis, Yarrowia lipolytica and Candida pseudoglaebosa, isolated from wastewater treatment plants, were tested for their ability to decolorize textile dyes. Both commercial textile synthetic dyes (reactive, disperse, direct, acid and basic) and simulated textile effluents (a total of 32 solutions) were added to a Normal Decolorization Medium along with the yeast (single strains and consortia) and the decolorization was evaluated spectrophotometrically for 48–72 h. Yeasts were able to perform decolorization through adsorption and biodegradation for 28 of the dyes and simulated effluents by more than 50%. Y. lipolytica and C. pseudoglaebosa presented the best results with a true decolorization of reactive dyes, above 90% at 100 mg l−1, and simulated effluents at 5 g l−1 of concentration. Enzyme production was evaluated: oxidoreductase was found in the three yeasts, whereas tyrosinase was only found in Y. lipolytica and C. pseudoglaebosa. Y. lipolytica and C. pseudoglaebosa are a potential biotechnological tool for dye degradation in textile wastewaters, especially those containing reactive dyes and a promising tool to integrate in bioremediation solutions, contributing to circular economy and eco sustainability in the water sector since the treated water could possibly be reused for irrigation.

Alginate-based microparticles coated with HPMCP/AS cellulose-derivatives enable the Ctx(Ile21)-Ha antimicrobial peptide application as a feed additive

Microencapsulation is a potential biotechnological tool, which can overcome antimicrobial peptides (AMP) instabilities and reduce toxic side effects. Thus, this study evaluates the antibacterial activities of the Ctx(Ile21)-Ha AMP against multidrug-resistant (MDR) and non-resistant bacteria and develop and characterize peptide-loaded microparticles coated with the enteric polymers hydroxypropylmethylcellulose acetate succinate (HPMCAS) and hydroxypropylmethylcellulose phthalate (HPMCP). Ctx(Ile21)-Ha was obtained by solid phase peptide synthesis (SPPS) method, purified and characterized by HPLC and Mass Spectrometry. The peptide exhibited potent antibiotic activities against Salmonella enteritidis, Salmonella typhimurium, Pseudomonas aeruginosa (MDR), Acinetobacter baumannii (MDR), and Staphylococcus aureus (MDR). Ctx(Ile21)-Ha microencapsulation was performed by ionic gelation with high efficiency, maintaining the physical-chemical stability. Ctx(Ile21)-Ha coated-microparticles were characterized by DSC, TGA, FTIR-Raman, XRD and SEM. Hemolytic activity assay demonstrated that hemolysis was decreased up to 95% compared to single molecule. In addition, in vitro release control profile simulating different portions of gastrointestinal tract was performed and showed the microcapsules' ability to protect the peptide and release it in the intestine, aiming pathogen's location, mainly by Salmonella sp. Therefore, use of microencapsulated Ctx(Ile21)-Ha can be allowed as an antimicrobial controller in monogastric animal production as an oral feed additive (antimicrobial controller), being a valuable option for molecules with low therapeutic indexes or high hemolytic rates.

Bioactive Metabolites from the Endophytic Fungus Aspergillus sp. SPH2.

In the current study, an ethyl acetate extract from the endophytic fungus Aspergillus sp. SPH2 isolated from the stem parts of the endemic plant Bethencourtia palmensis was screened for its biocontrol properties against plant pathogens (Fusarium moniliforme, Alternaria alternata, and Botrytis cinerea), insect pests (Spodoptera littoralis, Myzus persicae, Rhopalosiphum padi), plant parasites (Meloidogyne javanica), and ticks (Hyalomma lusitanicum). SPH2 gave extracts with strong fungicidal and ixodicidal effects at different fermentation times. The bioguided isolation of these extracts gave compounds 1–3. Mellein (1) showed strong ixodicidal effects and was also fungicidal. This is the first report on the ixodicidal effects of 1. Neoaspergillic acid (2) showed potent antifungal effects. Compound 2 appeared during the exponential phase of the fungal growth while neohydroxyaspergillic acid (3) appeared during the stationary phase, suggesting that 2 is the biosynthetic precursor of 3. The mycotoxin ochratoxin A was not detected under the fermentation conditions used in this work. Therefore, SPH2 could be a potential biotechnological tool for the production of ixodicidal extracts rich in mellein.

Engineering ribose-5-phosphate isomerase B from a central carbon metabolic enzyme to a promising sugar biocatalyst.

Ribose-5-phosphate isomerase B (RpiB) was first identified in the pentose phosphate pathway responsible for the inter-conversion of ribose-5-phosphate and ribulose-5-phosphate. Though there are seldom key enzymes in central carbon metabolic system developed as useful biocatalysts, RpiB with the advantages of wide substrate scope and high stereoselectivity has become a potential biotechnological tool to fulfill the demand of rare sugars currently. In this review, the pivotal roles of RpiB in carbon metabolism are summarized, and their sequence identity and structural similarity are discussed. Substrate binding and catalytic mechanisms are illustrated to provide solid foundations for enzyme engineering. Interesting differences in origin, physiological function, structure, and catalytic mechanism between RpiB and ribose-5-phosphate isomerase A are introduced. Moreover, enzyme engineering efforts for rare sugar production are stressed, and prospects of future development are concluded briefly in the viewpoint of biocatalysis. Aided by the progresses of structural and computational biology, the application of RpiB will be promoted greatly in the preparation of valuable molecules. KEY POINTS: • Detailed illustration of RpiB's vital function in central carbon metabolism. • Potential of RpiB in sequence, substrate scope, and mechanism for application. • Enzyme engineering efforts to promote RpiB in the preparation of rare sugars.

Crosstalk among hydrogen sulfide (H2S), nitric oxide (NO) and carbon monoxide (CO) in root-system development and its rhizosphere interactions: A gaseous interactome

Root development in higher plants is achieved by a precise intercellular communication which determines cell fate in the primary embryonic meristem where the gasotransmitters H2S, NO and CO participate dynamically. Furthermore, the rhizosphere interaction of these molecules with microbial and soil metabolism also affects root development. NO regulates root growth and architecture in association with several other biomolecules like auxin indole-3-acetic acid (IAA), ethylene, jasmonic acid (JA), strigolactones, alkamides and melatonin. The CO-mediated signal transduction pathway in roots is closely linked to the NO-mediated signal cascades. Interestingly, H2S acts also as an upstream component in IAA and NO-mediated crosstalk during root development. Heme oxygenase (HO) 1 generates CO and functions as a downstream component in H2S-mediated adventitious rooting and H2S–CO crosstalk. Likewise, reactive oxygen species (ROS), H2S and NO crosstalk are important components in the regulation of root architecture. Deciphering these interactions will be a potential biotechnological tool which could provide benefits in crop management in soils, especially under adverse environmental conditions. This review aims to provide a comprehensive update of the complex networks of these gasotransmitters during the development of roots.

Rhizophagus irregularis modulates cadmium uptake, metal transporter, and chelator gene expression in Medicago sativa.

Arbuscular mycorrhizal fungi (AMF) are considered a potential biotechnological tool for mitigating heavy metal (HM) toxicity. A greenhouse experiment was conducted to evaluate the impacts of the AM fungus Rhizophagus irregularis on cadmium (Cd) uptake, mycorrhizal colonization, and some plant growth parameters of Medicago sativa (alfalfa) in Cd-polluted soils. In addition, expression of two metal chelators (MsPCS1 (phytochelatin synthase) and MsMT2 (metallothionein)) and two metal transporter genes (MsIRT1 and MsNramp1) was analyzed using quantitative real-time PCR (qRT-PCR). Cd addition had a significant negative effect on mycorrhizal colonization. However, AMF symbiosis promoted the accumulation of biomass under both stressed and unstressed conditions compared with non-mycorrhizal (NM) plants. Results also showed that inoculation with R. irregularis significantly reduced shoot Cd concentration in polluted soils. Transcripts abundance of MsPCS1, MsMT2, MsIRT1, and MsNRAMP1 genes were downregulated compared with NM plants indicating that metal sequestration within hyphal fungi probably made Cd concentration insufficient in root cells for induction of these genes. These results suggest that reduction of shoot Cd concentration in M. sativa colonized by R. irregularis could be a promising strategy for safe production of this plant in Cd-polluted soils.

Effects of cryopreservation on mitochondrial function and sperm quality in fish

The development of cryopreservation techniques has led to important changes in animal reproductive biotechnology. However, these techniques are associated with cellular and molecular damage, affecting the mitochondrial function and quality of spermatozoa; moreover studies in fish are limited. In this work, the effects of cryopreservation on ultrastructure, mitochondrial function and antioxidant defences in Atlantic salmon (Salmo salar) spermatozoa were assessed, along with intracellular calcium (Ca2+)i, mitochondrial DNA sequence and sperm function (motility and fertilization rate). Significant ultrastructure alterations of the middle piece and mitochondria were observed in cryopreserved spermatozoa as compared to controls. Oxygen consumption and ATP were also significantly different in cryopreserved spermatozoa, and in spermatozoa incubated with electron transport chain (ETC) uncouplers and inhibitors. Mitochondrial membrane potential, motility, fertilization rate and Ca2+i in cryopreserved spermatozoa displayed significant reductions compared to fresh spermatozoa. Mitochondrial potential correlated significantly with motility and fertilization rate. A redox imbalance was observed in frozen spermatozoa due to increased lipid peroxidation and superoxide anion production as compared to fresh spermatozoa. Likewise, increased activity of glutathione peroxidase and total glutathione (GSH/GSSG), as well as reduced catalase activity, were observed in comparison with fresh spermatozoa. Our results contribute to a better understanding of cryodamage to mitochondrial functions in fish spermatozoa, and enabled us to establish potential quality assessment indicators. The data suggest that cryopreservation induces a reduction in overall sperm quality and functionality through disruption of the mitochondrial ultrastructure and function, leading to energy depletion and increased oxidative stress. This knowledge may also lead to the identification of a potential biotechnological tool for improving reproductive efficiency in species of commercial and biological interest.

Plant growth and their root development after inoculation of arbuscular mycorrhizal fungi in coal mine subsided areas

Coal mining often cause serious land degradation, soil erosion, and desertification affecting growth of the local vegetation, especially the roots. Arbuscular mycorrhizal fungi (AMF) inoculation is considered a potential biotechnological tool for mined soil remediation because mycorrhizal fungi could improve plant growth environment, especially under adverse conditions due to their good symbiosis. A field experiment was conducted to study the ecological effects of AMF (Funneliformis mosseae, Rhizophagus intraradices) on the growth of Amygdalus pedunculata Pall. and their root development in the regenerated mining subsidence sandy land. The reclamation experiment included four treatments: inoculation of Funneliformis mosseae (F.m), inoculation of Rhizophagus intraradices (R.i), combined inoculation of F.m and R.i and non-inoculated treatment. Root mycorrhizal colonization, plant height, crown width, soil moisture, root morphology and certain soil properties were assessed. The results showed that AMF improved the shoot and root growth of Amygdalus pedunculata Pall., and significantly increased root colonization after 1 year of inoculation. Available phosphorus content, activities of phosphatase as well as electrical conductivity in soil rhizosphere of all the three inoculation treatments were higher than that of the non-inoculated treatment. AMF increased the quantity of bacteria and fungi in soil rhizosphere compared with the non-inoculated treatment. Our study indicates that revegetation with AMF inoculum could influence plant growth and root development as well as soil properties, suggesting that AMF inoculation can be effective method for further ecological restoration in coal mine subsided areas.

Expression and characterization of a κ-carrageenase from marine bacterium Wenyingzhuangia aestuarii OF219: A biotechnological tool for the depolymerization of κ-carrageenan

κ-Carrageenases are desirable tools for tailoring the molecular weight, physicochemical properties and functionalities of κ-carrageenan, a macromolecule widely utilized in various industries. In this study, a novel GH16 family κ-carrageenase, designated as Cgk16A, was cloned from the genome of marine bacterium Wenyingzhuangia aestuarii OF219 and expressed in Escherichia coli. Its biochemical properties, kinetic parameters and hydrolytic pattern were characterized. The enzyme demonstrated a low optimal reaction temperature (25 °C) and a cold-adapted feature. As an endo-acting glycoside hydrolase, Cgk16A degraded κ-carrageenan in a random manner, and it was competent to prepare the degradation products with varying degrees of polymerization. The mass spectrometry analysis revealed that the end products of Cgk16A were majorly composed of κ-carrageenan tetrasaccharide with a minor portion of disaccharide. The enzyme showed higher enzyme-substrate affinity over all hitherto characterized GH16 κ-carrageenases, indicated by its low Km value (0.17 μM). Cgk16A could be employed as a potential biotechnological tool for depolymerizing κ-carrageenan, which would facilitate the future application of κ-carrageenan and its degradation products.

Potential of different AM fungi (native from As-contaminated and uncontaminated soils) for supporting Leucaena leucocephala growth in As-contaminated soil

Arbuscular mycorrhizal (AM) fungi inoculation is considered a potential biotechnological tool for an eco-friendly remediation of hazardous contaminants. However, the mechanisms explaining how AM fungi attenuate the phytotoxicity of metal(oid)s, in particular arsenic (As), are still not fully understood. The influence of As on plant growth and the antioxidant system was studied in Leucaena leucocephala plants inoculated with different isolates of AM fungi and exposed to increasing concentrations of As (0, 35, and 75 mg dm−3) in a Typic Quartzipsamment soil. The study was conducted under greenhouse conditions using isolates of AM fungi selected from uncontaminated soils (Acaulospora morrowiae, Rhizophagus clarus, Gigaspora albida; and a mixed inoculum derived from combining these isolates, named AMF Mix) as well as a mix of three isolates from an As-contaminated soil (A. morrowiae, R. clarus, and Paraglomus occultum). After 21 weeks, the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) were determined in the shoots in addition to measuring plant height and mineral contents. In general, AM fungi have shown multiple beneficial effects on L. leucocephala growth. Although the activity of most of the stress-related enzymes increased in plants associated with AM fungi, the percentage increase caused by adding As to the soil was even greater for non-mycorrhizal plants when compared to AM-fungi inoculated ones, which highlights the phytoprotective effect provided by the AM symbiosis. The highest P/As ratio observed in AM-fungi plants, compared to non-mycorrhizal ones, can be considered a good indicator that the AM fungi alter the pattern of As(V) uptake from As-contaminated soil. Our results underline the role of AM fungi in increasing the tolerance of L. leucocephala to As stress and emphasize the potential of the symbiosis L. leucocephala-R. clarus for As-phytostabilization at moderately As-contaminated soils.

Phytoremediation potential of rhizobacterial isolates from Kans grass (Saccharum spontaneum) of fly ash ponds

Kans grass (Saccharum spontaneum) is a weed species that is frequently found in many heavy metal-enriched waste dumps including fly ash pond sites. In this study, among a collection of phosphate-solubilizing bacterial strains isolated from the rhizosphere of Saccharum spontaneum present in the abandoned ash pond site of Mejia Thermal Power Station (MTPS-DVC), three strains were characterized for their plant growth-promoting abilities. The isolates identified as Bacillus anthracis strain MHR2, Staphylococcus sp. strain MHR3 and Bacillus sp. strain MHR4 had phosphate solubilization indices of 2.86, 2.31 and 2.40 and they produced soluble phosphates of 700, 600 and 640 mg l−1, respectively, in 4 days. In all the PSBs, pH significantly decreased, indicating the production of various organic acids. They showed other plant growth-promoting features like production of ammonia, siderophore, hydrocyanide and IAA. All of them were resistant to multiple heavy metals and antibiotics. Dry and fresh weight and shoot and root lengths of Brassica juncea L. increased in the presence of these isolates in pot cultures. The strains also increased phytoextraction ability of plants by enhancing the metal accumulation in plant tissues. Thus, the isolated indigenous and stress-adapted rhizobacteria may serve as potential biotechnological tool for the successful ecorestoration of various metal-contaminated sites.

Epoxide hydrolase of Trichoderma reesei: Biochemical properties and conformational characterization

Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have biotechnological potential in chiral chemistry. We report the cloning, purification, enzymatic activity, and conformational analysis of the TrEH gene from Trichoderma reesei strain QM9414 using circular dichroism spectroscopy. The EH gene has an open reading frame encoding a protein of 343 amino acid residues, resulting in a molecular mass of 38.2kDa. The enzyme presents an optimum pH of 7.2, and it is highly active at temperatures ranging from 23 to 50°C and thermally inactivated at 70°C (t1/2=7.4min). The Michaelis constants (Km) were 4.6mM for racemic substrate, 21.7mM for (R)-(+)-styrene oxide and 3.0mM for (S)-(−)-styrene oxide. The kcat/Km analysis indicated that TrEH is enantioselective and preferentially hydrolyzes (S)-(−)-styrene oxide. The conformational stability studies suggested that, despite the extreme conditions (high temperatures and extremely acid and basic pHs), TrEH is able to maintain a considerable part of its regular structures, including the preservation of the native cores in some cases. The recombinant protein showed enantioselectivity that was distinct from other fungus EHs, making this protein a potential biotechnological tool.

The Combined Effects of Arbuscular Mycorrhizal Fungi (AMF) and Lead (Pb) Stress on Pb Accumulation, Plant Growth Parameters, Photosynthesis, and Antioxidant Enzymes in Robinia pseudoacacia L.

Arbuscular mycorrhizal fungi (AMF) are considered as a potential biotechnological tool for improving phytostabilization efficiency and plant tolerance to heavy metal-contaminated soils. However, the mechanisms through which AMF help to alleviate metal toxicity in plants are still poorly understood. A greenhouse experiment was conducted to evaluate the effects of two AMF species (Funneliformis mosseae and Rhizophagus intraradices) on the growth, Pb accumulation, photosynthesis and antioxidant enzyme activities of a leguminous tree (Robinia pseudoacacia L.) at Pb addition levels of 0, 500, 1000 and 2000 mg kg-1 soil. AMF symbiosis decreased Pb concentrations in the leaves and promoted the accumulation of biomass as well as photosynthetic pigment contents. Mycorrhizal plants had higher gas exchange capacity, non-photochemistry efficiency, and photochemistry efficiency compared with non-mycorrhizal plants. The enzymatic activities of superoxide dismutase (SOD), ascorbate peroxidases (APX) and glutathione peroxidase (GPX) were enhanced, and hydrogen peroxide (H2O2) and malondialdehyde (MDA) contents were reduced in mycorrhizal plants. These findings suggested that AMF symbiosis could protect plants by alleviating cellular oxidative damage in response to Pb stress. Furthermore, mycorrhizal dependency on plants increased with increasing Pb stress levels, indicating that AMF inoculation likely played a more important role in plant Pb tolerance in heavily contaminated soils. Overall, both F. mosseae and R. intraradices were able to maintain efficient symbiosis with R. pseudoacacia in Pb polluted soils. AMF symbiosis can improve photosynthesis and reactive oxygen species (ROS) scavenging capabilities and decrease Pb concentrations in leaves to alleviate Pb toxicity in R. pseudoacacia. Our results suggest that the application of the two AMF species associated with R. pseudoacacia could be a promising strategy for enhancing the phytostabilization efficiency of Pb contaminated soils.

92 - Nitroxyl as a Positive Inotropic Therapy in Heart Failure: cMyBP-C as a Target?

Root development in higher plants is achieved by a precise intercellular communication which determines cell fate in the primary embryonic meristem where the gasotransmitters H2S, NO and CO participate dynamically. Furthermore, the rhizosphere interaction of these molecules with microbial and soil metabolism also affects root development. NO regulates root growth and architecture in association with several other biomolecules like auxin indole-3-acetic acid (IAA), ethylene, jasmonic acid (JA), strigolactones, alkamides and melatonin. The CO-mediated signal transduction pathway in roots is closely linked to the NO-mediated signal cascades. Interestingly, H2S acts also as an upstream component in IAA and NO-mediated crosstalk during root development. Heme oxygenase (HO) 1 generates CO and functions as a downstream component in H2S-mediated adventitious rooting and H2S–CO crosstalk. Likewise, reactive oxygen species (ROS), H2S and NO crosstalk are important components in the regulation of root architecture. Deciphering these interactions will be a potential biotechnological tool which could provide benefits in crop management in soils, especially under adverse environmental conditions. This review aims to provide a comprehensive update of the complex networks of these gasotransmitters during the development of roots.

The sunflower transcription factor HaWRKY76 confers drought and flood tolerance to Arabidopsis thaliana plants without yield penalty.

Arabidopsis transgenic plants expressing the sunflower transcription factor HaWRKY76 exhibit increased yield and tolerance to drought and flood stresses. The genetic construct containing HaWRKY76 is proposed as a potential biotechnological tool to improve crops. Water deficit and water excess are abiotic stress factors that seriously affect crops worldwide. To increase the tolerance to such stresses without causing yield penalty constitutes a major goal for biotechnologists. In this survey, we report that HaWRKY76, a divergent sunflower WRKY transcription factor, is able to confer both dehydration and submergence tolerance to Arabidopsis transgenic plants without yield penalty. The expression pattern of HaWRKY76 was analyzed in plants grown in standard conditions and under different watering regimes indicating a regulation by water availability. The corresponding cDNA was isolated and cloned under the control of a constitutive promoter and Arabidopsis plants were transformed with this construct. These transgenic plants presented higher biomass, seed production and sucrose content than controls in standard growth conditions. Moreover, they exhibited tolerance to mild drought or flood (complete submergence/waterlogging) stresses as well as the same or increased yield, depending on the stress severity and plant developmental stage, compared with controls. Drought tolerance occurred via an ABA-independent mechanism and induction of stomatal closure. Submergence tolerance can be explained by the carbohydrate (sucrose and starch) preservation achieved through the repression of fermentation pathways. Higher cell membrane stability and chlorenchyma maintenance could be the nexus between tolerance responses in front of both stresses. Altogether, the obtained results indicated that HaWRKY76 can be a potential biotechnological tool to improve crops yield as well as drought and flood tolerances.

Antibacterial activity of the Antarctic bacterium Janthinobacterium sp. SMN 33.6 against multi-resistant Gram-negative bacteria

article i nfo Background: The increment of resistant strains to commonly used antibiotics in clinical practices places in evidencetheurgentneedtosearchfornewcompoundswith antibacterial activity.TheadaptationsthatAntarctic microorganismshavedeveloped,due tothe extremeenvironmentthattheyinhabit, promote themasa potential new source of active compounds for the control of microorganisms causing infections associated with health care.TheaimofthisstudywastoevaluatetheantibacterialactivityofanethanolextractoftheAntarcticbacterium Janthinobacterium sp., strain SMN 33.6, against nosocomial multi-resistant Gram-negative bacteria. Results: Inhibitory activity against human Gram-negative bacterial pathogens, with concentrations that varied between 0.5 and 16 μ gm l -1 , was demonstrated. Conclusions:The ethanolic extract of Janthinobacterium sp. SMN 33.6 possesses antibacterial activity against a chromosomal AmpC beta-lactamase-producing strain of Serratia marcescens, an extended-spectrum beta-lactamase-producing Escherichia coli and also against carbapenemase-producing strains of Acinetobacter baumannii and Pseudomonas aeruginosa. This becomes a potential and interesting biotechnological tool for the control of bacteria with multi-resistance to commonly used antibiotics.